Cell Signaling Technology

Product Pathways - Neuroscience

Phospho-CREB (Ser133) (D1G6) Rabbit mAb #4276

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP ChIP H M R Endogenous 43 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-CREB (Ser133) (D1G6) Rabbit mAb detects endogenous levels of CREB only when phosphorylated at Ser133. The antibody also detects the phosphorylated form of the CREB-related protein, ATF-1.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser133 of human CREB protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from SH-SY5Y cells, untreated or treated with Forskolin #3828, using Phospho-CREB (Ser133) (D1G6) Rabbit mAb (upper) or CREB (D76D11) Rabbit mAb #4820 (lower).

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 293 cells treated with Forskolin #3828 (30 µM) for 1 hour and either 10 μl of Phospho-CREB (Ser133) (D1G6) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human ALS2 exon 1 primers, SimpleChIP® Human NR4A3 Promoter Primers #4829, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Background

CREB is a bZIP transcription factor that activates target genes through cAMP response elements. CREB is able to mediate signals from numerous physiological stimuli, resulting in regulation of a broad array of cellular responses. While CREB is expressed in numerous tissues, it plays a large regulatory role in the nervous system. CREB is believed to play a key role in promoting neuronal survival, precursor proliferation, neurite outgrowth, and neuronal differentiation in certain neuronal populations (1-3). Additionally, CREB signaling is involved in learning and memory in several organisms (4-6). CREB is able to selectively activate numerous downstream genes through interactions with different dimerization partners. CREB is activated by phosphorylation at Ser133 by various signaling pathways including Erk, Ca2+, and stress signaling. Some of the kinases involved in phosphorylating CREB at Ser133 are p90RSK, MSK, CaMKIV, and MAPKAPK-2 (7-9).

  1. Lonze, B.E. et al. (2002) Neuron 34, 371-85.
  2. Lee, M.M. et al. (1999) J Neurosci Res 55, 702-12.
  3. Redmond, L. et al. (2002) Neuron 34, 999-1010.
  4. Dash, P.K. et al. (1990) Nature 345, 718-21.
  5. Yin, J.C. et al. (1994) Cell 79, 49-58.
  6. Guzowski, J.F. and McGaugh, J.L. (1997) Proc Natl Acad Sci USA 94, 2693-8.
  7. Xing, J. et al. (1998) Mol Cell Biol 18, 1946-55.
  8. Ribar, T.J. et al. (2000) J Neurosci 20, RC107.
  9. Tan, Y. et al. (1996) EMBO J 15, 4629-42.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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